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OALib Journal期刊
ISSN: 2333-9721
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黄嘌呤氧化酶抑制剂/超氧阴离子清除剂双靶点高通量筛选模型的建立
, PP. 447-452
Keywords: 高通量筛选方法,黄嘌呤氧化酶,超氧阴离子清除剂,抗氧化,药物评价
Abstract:
本文建立了适用于同时筛选黄嘌呤氧化酶抑制剂和超氧阴离子清除剂的双靶点高通量复合筛选模型.在黄嘌呤氧化酶超氧阴离子生成体系中,加入wst-1作为超氧阴离子生成量的探针,以反应体系中标识性产物尿酸为黄嘌呤氧化酶活性指示剂,采用spectramaxm5酶标测试仪,同时检测两种指示剂的浓度变化,通过对反应体系中的影响因素进行优化建立双靶点hts筛选模型,并利用阳性药物对该模型进行评价.在反应体系中,反应终体积50μl,黄嘌呤氧化酶4mu·ml-1、黄嘌呤250μmol·l-1、wst-1浓度为100μmol·l-1,黄嘌呤氧化酶抑制剂筛选模型的z'-因子为0.5374,s/n为47.5199;超氧阴离子清除剂筛选模型的z'-因子为0.5074,s/n为5.3889.结果表明,本文建立的黄嘌呤氧化酶抑制剂/氧自由基清除剂高通量筛选模型具有稳定性好、成本较低和重复性高等特点,可以广泛应用于药物筛选.
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